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EPIDERMAL GROWTH FACTOR RECEPTOR (EGFR) PROTEIN SRM/MRM ASSAY

机译:表皮生长因子受体(EGFR)蛋白SRM / MRM分析

摘要

The current disclosure provides for specific peptides, and derived ionization characteristics of the peptides, from the Epidermal Growth Factor Receptor (EGFR) protein that are particularly advantageous for quantifying the EGFR protein directly in biological samples that have been fixed in formalin by the method of Selected Reaction Monitoring (SRM) mass spectrometry, or what can also be termed as Multiple Reaction Monitoring (MRM) mass spectrometry. Such biological samples are chemically preserved and fixed wherein said biological sample is selected from tissues and cells treated with formaldehyde containing agents/fixatives including formalin-fixed tissue/cells, formalin-fixed/paraffin embedded (FFPE) tissue/cells, FFPE tissue blocks and cells from those blocks, and tissue culture cells that have been formalin fixed and or paraffin embedded. A protein sample is prepared from said biological sample using the Liquid Tissue reagents and protocol and the EGFR protein is quantitated in the Liquid Tissue sample by the method of SRM/MRM mass spectrometry by quantitating in the protein sample at least one or more of the peptides described. These peptides can be quantitated if they reside in a modified or an unmodified form. An example of a modified form of an EGFR peptide is phosphorylation of a tyrosine, threonine, serine, and/or other amino acid residues within the peptide sequence.
机译:本公开提供了表皮生长因子受体(EGFR)蛋白的特异性肽和衍生的肽的电离特性,这些蛋白对于直接定量已经通过福尔马林固定在福尔马林中的生物样品中的EGFR蛋白特别有利。反应监测(SRM)质谱,或也可以称为多反应监测(MRM)质谱。这样的生物样品被化学保存并固定,其中所述生物样品选自用含甲醛的试剂/固定剂处理的组织和细胞,包括甲醛固定的组织/细胞,甲醛固定/石蜡包埋的(FFPE)组织/细胞,FFPE组织块和那些块中的细胞,以及已被福尔马林固定和/或石蜡包埋的组织培养细胞。使用液体组织试剂和规程从所述生物样品中制备蛋白质样品,并通过SRM / MRM质谱法通过在蛋白质样品中定量至少一种或多种肽来定量液体组织样品中的EGFR蛋白。描述。如果这些肽以修饰或未修饰的形式存在,则可以定量。 EGFR肽的修饰形式的实例是肽序列内的酪氨酸,苏氨酸,丝氨酸和/或其他氨基酸残基的磷酸化。

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